Susceptibility of Human Sperm toin SituDNA Denaturation Is Strongly Correlated with DNA Strand Breaks Identified by Single-Cell Electrophoresis
| Autor: | JA Bjordahl, G.R. Aravindan, Lorna K. Jost, Donald P. Evenson |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
Electrophoresis
Male Hot Temperature Biotin Semen DNA Fragmentation Alkalies Biology chemistry.chemical_compound Humans Denaturation (biochemistry) Cobalt Radioisotopes Staining and Labeling Dose-Response Relationship Radiation Cell Biology Hydrogen-Ion Concentration Flow Cytometry Spermatozoa Sperm Molecular biology Deoxyuridine Chromatin Comet assay chemistry DNA fragmentation Deoxyuracil Nucleotides Acids DNA |
| Zdroj: | Experimental Cell Research. 236:231-237 |
| ISSN: | 0014-4827 |
| Popis: | Susceptibility of mammalian sperm DNA to low pH- or heat-induced denaturation in situ has shown very strong dose-response relationships with animal and human exposure to chemical and physical toxicants and also fertility potential. In this study, 23 human semen samples representing a wide range in percentage (7-86%) of sperm exhibiting abnormally high susceptibility of DNA in situ to denaturation were studied for the integrity of their DNA using alkaline comet assay (single-cell microgel electrophoresis, pH 10.0). The percentage of comets observed for these samples ranged from 5 to 95%; these data correlated strongly with the percentage of sperm with increased DNA denaturability (r = 0.973; P < 0.001). Labeling of 3' ends of nicked DNA sites with 5-bromo-2'-deoxyuridine 5'-triphosphate (BrdUTP) followed by tagging with FITC-BrdUTP monoclonal antibody and flow cytometry also indicated significantly strong correlations of BrdUTP incorporation with both abnormal susceptibility of DNA to denaturation (r = 0.859, P < 0.001) and comet assay (r = 0.812, P < 0.001). The relationship among susceptibility of sperm chromatin to acid denaturation in situ, BrdUTP incorporation, and formation of comets suggests that DNA fragmentation monitored by these assays may have important physiological relevance in terms of sperm quality and fertility potential. |
| Databáze: | OpenAIRE |
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